A revised protocol for in vitro development of mouse oocytes from primordial follicles dramatically improves their developmental competence
A revised protocol for in vitro development of mouse oocytes from primordial follicles dramatically improves their developmental competence
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DOI：
10.1095/biolreprod.102.013029
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发表时间：
2003-05-01
影响因子：
3.6
通讯作者：
Eppig, JJ
中科院分区：
文献类型：
--
作者：
O'Brien, MJ;Pendola, JK;Eppig, JJ
本研究的目的是从新生小鼠的原始卵泡开始，改善卵母细胞体外发育的条件。以往的研究表明，具有减数分裂成熟、受精和植入前能力的卵母细胞可以从原始卵泡体外发育而来。然而，成功率很低，只有一个活的后代（0.5%的胚胎移植）。以卵母细胞成熟和着床前发育为终点，将修订后的方案与原始方案进行比较。使用修订后的方案，卵母细胞成熟至中期11和发育至囊胚期的百分比显著提高。此外，我们比较了来自体外生长和体内生长的卵母细胞的两细胞期胚胎的后代生产。在1160个移植的来自体外培养的卵母细胞的二细胞期胚胎中，66个（5.7%）发育至足月，7个幼崽（10.6%）在出生时死亡。其余59只幼崽（27只雌性，32只雄性）存活至成年。相比之下，在437个来自体内生长的卵母细胞的移植的两细胞期胚胎中，76个（17.4%）发育到足月，4个（5.3%）在出生时死亡。其余72只幼仔（35只雌性，37只雄性）存活至成年。这些研究为哺乳动物原始卵泡体外发育成完全有能力的卵母细胞提供了证据，并在卵母细胞培养技术方面取得了重大进展。
The objective of this study was to improve the conditions for oocyte development in vitro beginning with the primordial follicles of newborn mice. Previous studies showed that oocytes competent of meiotic maturation, fertilization, and preimplantation could develop in vitro from primordial follicles. However, the success rates were low and only one live offspring was produced (0.5% of embryos transferred). A revised protocol was compared with the original protocol using oocyte maturation and preimplantation development as end points. The percentage of oocytes maturing to metaphase 11 and developing to the blastocyst stage was significantly improved using the revised protocol. In addition, we compared the production of offspring from two-cell stage embryos derived from in vitro-grown and in vivo-grown oocytes. Of 1160 transferred two-cell stage embryos derived from in vitro-grown oocytes, 66 (5.7%) developed to term and 7 pups (10.6%) died at birth. The remaining 59 pups (27 females, 32 males) survived to adulthood. By comparison, of 437 transferred two-cell stage embryos derived from in vivo-grown oocytes, 76 (17.4%) developed to term and 4 (5.3%) died at birth. The remaining 72 pups (35 females, 37 males) survived to adulthood. These studies provide proof of the principle that fully competent mammalian oocytes can develop in vitro from primordial follicles and present a significant advance in oocyte culture technology.